Additional file 4.
PCR conditions for 11 microsatellite loci for A. germinans (black mangrove) [[65,66]] and six microsatellite loci for R. mangle (red mangrove) [] following established protocols [[13,65,66]] with modifications including three primers in the PCR thus: a dye tagged M13 universal forward primer (5’-CACGACGTTGTAAAACGAC-3’)[], primer Forward (F) and primer Reverse (R) where either F or R primer (indicated with asterisk *) had a tail at the 5’ end that was identical to the M13 universal forward primer sequence. Amplified fragments from both mangrove species were electrophoretically separated on ABI 3130xl and analyzed using ABI PRISM® GeneMapper™ software version 3.7. For each estuary is indicated the frequency of null alleles calculated by the Brookfield method 1 in MICROCHECKER 2.2.3 .
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Cerón-Souza et al. BMC Evolutionary Biology 2012 12:205 doi:10.1186/1471-2148-12-205